The CASP3 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population from Raji B lymphocytes with disrupted CASP3 gene. This model removes a central apoptosis effector, aiding dissection of cell death mechanisms in Burkitt’s lymphoma. The polyclonal format retains genetic diversity while ensuring target loss-of-function, suitable for non-clonal studies. The CRISPR disruption is marker-free, maintaining cellular physiology, and provides a ready-to-use tool for functional genomics and drug discovery.
The Raji cell line, from a Burkitt’s lymphoma patient, is an EBV-positive B lymphocyte model for B cell biology, lymphomagenesis, and EBV pathologies. Raji cells express B cell markers (CD19, CD20) and key lymphoma survival pathways. Their rapid growth and suspension culture facilitate high-throughput screening. As an aggressive lymphoma model, Raji cells offer a clinically relevant system for apoptosis resistance and B cell-targeted therapy studies.
Caspase-3 is the principal executioner caspase, integrating extrinsic death receptor and intrinsic mitochondrial apoptotic signals. It is activated by initiator caspase-8 and caspase-9, downstream of TNF, FasL, or mitochondrial cytochrome c release regulated by Bid and Bax. Active caspase-3 cleaves substrates like PARP, ICAD/DFF45, DFF40/CAD, gelsolin, and fodrin, causing DNA fragmentation and cellular demolition. IAPs (XIAP, survivin, cIAP1/2) inhibit caspase-3, while chaperones HSP70 and Hsp90 modulate signaling. In the apoptosome, caspase-3 functions downstream of Apaf-1 and cytochrome c. CASP3 disruption in Raji cells eliminates a critical node in the caspase cascade.
In Raji lymphoma cells, CASP3 knockout allows exploration of apoptosis-independent functions and adaptive responses to impaired cell death. Given the Burkitt’s lymphoma origin, it enables study of lymphomagenesis sustained without executioner caspase activity, potentially revealing alternative death pathways. The model is apt for investigating EBV latency and apoptotic regulation. CASP3-deficient Raji cells also serve for functional complementation to identify caspase-3 substrates and interactors in B cells.
Researchers use these cells in Annexin V apoptosis assays, TUNEL for DNA fragmentation, Western blotting for cleaved PARP and active caspase-3, and flow cytometry for caspase activity. Drug sensitivity profiling assesses caspase-3-dependent cytotoxicity. Applications include B cell development studies, complementation with CASP3 constructs, and CRISPR screens for apoptosis-resistant dependencies. For further information, contact Ascent Research.
